Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion

A new beta-lactam-inducible penicillin-binding protein (PBP) that has extremely low affinity to penicillin and most other beta-lactam antibiotics has been widely found in highly beta-lactam(methicillin)-resistant Staphylococcus aureus (MRSA). The gene for this protein was sequenced and the nucleotide sequence in its promoter and close upstream area was found to show close similarity with that of staphylococcal penicillinase, while the amino acid sequence over a wide range of the molecule was found to be similar to those of two PBPs of Escherichia coli, the shape-determining protein (PBP 2) and septum-forming one (PBP 3). Probably the MRSA PBP (Mr 76462) evolved by recombination of two genes: an inducible type I penicillinase gene and a PBP gene of a bacterium, causing the formation of a beta-lactam-inducible MRSA PBP.     

稻田蜘蛛混合种群空间分布模式动态分析(英文)

结果表明,在东方红农场稻田中总共有11科29属43种蜘蛛,其中早稻田中有8科19属43种,晚稻田中有10科27属36种。在水稻的不同发育阶段蜘蛛混合种群的空间分布模式存在差异。在早稻发育的前期、中期和后期,稻田蜘蛛混合种群的空间分布模式分别是聚集的、随机的和聚集的;在晚稻发育的前期、中期和后期,其空间分布模式依次是均匀的、聚集的和均匀的。同时,对其空间分布模式动态进行了初步分析。     

感染羊瘙痒因子263K株或139A株仓鼠脑组织中tau蛋白和磷酸化tau蛋白变化的研究

为探讨朊病毒病的神经病理特征,应用Western blot方法检测了感染羊瘙痒因子263K株或139A株的仓鼠脑组织中总tau蛋白和Ser396和Ser404位点发生磷酸化tau蛋白表达水平的变化;并应用Real Time PCR方法检测了tau mRNAs转录活性的改变。结果表明总tau蛋白含量升高而Ser396和Ser404位点发生磷酸化的tau蛋白含量降低,该现象与羊瘙痒因子毒株类型和临床潜伏期无关;感染羊瘙痒因子的仓鼠脑组织中Tau2和Tau4这两个异构体的转录水平升高。这些结果表明tau蛋白在Ser396和Ser404位点的去磷酸化可能与朊病毒病发病相关。     

家蝇幼虫血淋巴中抗真菌肽的诱导方法比较及抗真菌活性

以未诱导组作为空白对照研究比较真菌诱导、超声诱导和热诱导家蝇Musca domestica 幼虫血淋巴初提液的抗真菌肽效果,比较各种诱导方法诱导后的幼虫存活率;用凝胶层析法和高效液相分离纯化热诱导家蝇3龄幼虫抗真菌肽,检测其抗白假丝酵母菌Candida albicans和新生隐球菌Cryptococcus neoformans活性;SDS-PAGE分析抗真菌肽的蛋白分子量范围。结果表明：3种诱导方法诱导后家蝇幼虫均产生具有明显抗真菌作用的抗真菌肽,其初提液抑菌圈大小没有明显差别;真菌诱导组和热诱导组幼虫存活率低于对照组,而超声诱导组与对照组相比则无明显差别。经分离纯化后,抗真菌肽仍具有较好的抗真菌活性;SDS-PAGE分析表明该抗真菌肽有效成分的蛋白分子量在14.4 kD以下。结果提示热诱导家蝇幼虫产生抗真菌肽是一种方便、有效的诱导方式。     

禽病原性大肠杆菌1型菌毛的分离与鉴定

以旋涡混合法使禽病原性大肠杆菌分离株566、1794和TK3菌毛脱落,经硫酸铵沉淀、透析后进行蔗糖密度梯度离心,收集密度为110至115g/cm3的蛋白带,经SDSPAGE测定,3株菌菌毛蛋白的分子量分别在175、170和170kD;提纯菌毛保留了甘露糖敏感性凝集豚鼠红细胞的能力,证明它们为1型菌毛;从1794株提取的1型菌毛免疫BALB/C小鼠产生的高免血清在Western blot中与3个菌株的相应菌毛蛋白均呈阳性反应。上述结果表明,受检的3株禽病原性大肠杆菌均表达了1型菌毛,其分子量在175～170kD之间,3个菌株的1型菌毛间具有较强的抗原相关性。     

Homology modeling threedimensional structure of AnxB1 and reducing its immunogenicity by sequence-deleted mutagenesis

AnxB1,a novel annexin previously isolated from Cysticercus cellulose,shows high thrombi affinity and anticoagulant activity in vivo.In order to investigate the relationship between structure and biological function,a predicted three-dimensional(3D)model of AnxB1 was generated by homology modeling.This model contains four homologous internal-domains and the Cα trace of domain Ⅰ,Ⅱ and IV shows high similarity.Based on the structure characterization,four sequence-deleted mutants were constructed and expressed as GST fusion proteins in E.coli.Two of the mutants,GST-M3 and GST-M4 reserved high anticoagulant activity(p＜0.01 vs.GST).Furthermore,compared with the wild type GST-AnxB1,the immunogenicity of GST-M3 and GST-M4 was reduced significantly(p＜0.01)and the molecular weight was lowered to 27 kD and 34 kD,respectively.These observations laid a solid foundation for further study on developing new thrombolytic agents with higher efficiency and lower side effect.     

UreB酸奶的研制及其抗幽门螺杆菌感染的动物实验研究

目的 利用UreB基因工程乳杆菌(UreB-L6032)研制一种预防幽门螺杆菌(Helicobacter pylori,H.pylori)感染的UreB活性酸奶,并对其抗H.pylori感染的能力进行动物实验研究.方法 采用发酵奶制备技术制成酸奶,并对其pH、活菌含量和质粒稳定性进行检测.小鼠在喂食UreB活性酸奶1个月后再感染H.pylori,并设普通酸奶组,阴性和阳性感染组,2个月后测定IL-10(interleukin-10)、IFN-γ(interferon-γ)、sIgA和特异性IgG的水平,并利用快速脲酶法和细菌培养法对小鼠胃部H.pylori进行检测.结果 成功的研制出了色香味颇佳,含UreB-L6032活菌较高,质粒能稳定存在的UreB活性酸奶.与一般酸奶相比,其能诱导小鼠体内特异性IgG的产生,显著增加IL-10的表达,降低小鼠H.pylori感染率.结论 UreB活性酸奶比一般酸奶能更加有效预防H.pylori感染.     

Purification and spectroscopic properties of 124-kDa oat phytochrome

A simplified procedure for the isolation and purification of 124-kDa phytochrome from etiolated Avena seedlings has been developed using the method of ammonium sulfate back-extraction. After hydroxyapatite chromatography of seedling tissue extracts, the pooled phytochrome was subjected to ammonium sulfate back-extraction instead of the usual application to an Affi-Gel Blue column. The resulting phytochrome had specific absorbance ratios (SAR = A666/A280) ranging from 0.85 to 0.95. Subsequent Bio-Gel filtration chromatography yielded highly pure 124-kDa phytochrome with SAR values ranging from 0.99 to 1.13. The absorption maxima of 124-kDa phytochrome were at 280, 379, and 666 nm for the red absorbing form of phytochrome (Pr) and at 280, 400 and 730 nm for the far-red absorbing form (Pfr). The A730/A673 ratio in Pfr was found to be 1.5 to 1.6. The mole fraction of Pfr under red light photoequilibrium was 0.88. No dark reversion was detected within 5 h at 3 degrees C. A photoreversible far-uv-circular dichroism was observable with all phytochrome preparations examined. Fluorescence and phosphorescence lifetimes were measured to further characterize the differences between the phytochromes prepared under different conditions. The Trp fluorescence and phosphorescence lifetimes of Pr and Pfr with the chromophore "X", probably polyphenolic in nature, were significantly shorter than those of phytochrome without the contaminant X. The short lifetime of the fluorescence of the Pr chromophore is attributable to X in the former.     

Seasonal occurrence of helminths in the anadromous fish Coilia nasus (Engraulidae): parasite indicators of fish migratory movements

To understand the seasonal migration of the anadromous Coilia nasus , we attempted to identify the parasites infecting C. nasus and determine their seasonal occurrence. From June 2007 to July 2008, a survey of 775 C. nasus individuals from the estuary of the Yangtze River and the coast of the East China Sea revealed more than 7,300 parasites associated with the gills and alimentary tracts of C. nasus . The following 6 helminth taxa were identified, i.e., the monogeneans Heteromazocraes lingmueni and Helciferus tenuis, the digenean Elytrophallus coiliae, the acanthocephalan Acanthosentis cheni , and larvae of the nematodes Anisakis simplex and Contracaecum sp., all of which are marine or brackish-water parasites. The absence of freshwater helminths suggested that the parasites acquired in freshwater may have been accidentally, and easily, lost by the time the fish had reached the estuary and coast. Contrary to seasonal occurrence of the parasites' life cycles, the lowest mean abundance and prevalence of H. lingmueni and A. cheni occurred in August, which suggested the immigration of C. nasus from freshwater to the Yangtze estuary, with lower parasite burdens. The highest mean abundance and prevalence of the nematodes A. simplex and Contracaecum sp. in May and June, and the lowest in August, indicated the arrival of the fish from the coast and the Yangtze River, to the estuary, respectively. These findings suggested that a majority of the fish prepared for spawning migration in the estuary in spring and early summer and returned to the estuary after spawning in the lower and middle reaches of the Yangtze River in late summer.